本文選題：阿勒泰羊 + 饑餓與非饑餓模型��； 參考：《石河子大學(xué)》2015年碩士論文

【摘要】：目的:阿勒泰羊是一種脂臀型綿羊品種,其獨(dú)特的尾脂在維持機(jī)體正常的新陳代謝和體溫過(guò)程中扮演了重要的角色,主要通過(guò)尾部脂肪沉積與代謝消長(zhǎng)規(guī)律來(lái)適應(yīng)惡劣的環(huán)境以維持機(jī)體正常的新陳代謝和體溫,是研究綿羊尾脂沉積與代謝機(jī)制的理想模型。近年來(lái)的研究表明,FABP4、ADIPOQ和CFD基因參與葡萄糖和脂肪酸代謝以及脂肪細(xì)胞分化等生理過(guò)程,初步推測(cè)其與脂肪沉積代謝密切相關(guān)。本文以阿勒泰羊?yàn)檠芯繉?duì)象,探討了FABP4、ADIPOQ、CFD基因?qū)Π⒗仗┭蛭仓练e與代謝的影響。方法:1、阿勒泰羊饑餓與非饑餓模型的構(gòu)建。模擬阿勒泰羊在自然生理狀態(tài)下尾脂沉積與代謝兩種極端狀態(tài),即在夏秋季節(jié)牧草豐茂時(shí)脂肪沉積、春冬季節(jié)牧草匱乏時(shí)尾脂分解代謝的方式,對(duì)阿勒泰羊進(jìn)行了饑餓試驗(yàn),構(gòu)建了阿勒泰羊饑餓與非饑餓狀態(tài)下尾脂沉積與代謝的動(dòng)物模型。2、以阿勒泰羊尾脂組織c DNA為模板,通過(guò)RT-PCR方法克隆獲得FABP4、ADIPOQ和CFD基因完整的CDS區(qū)序列。3、以阿勒泰羊心臟、肝臟、脾臟、肺臟、腎臟、肌肉、腸系膜脂肪、尾脂等主要組織c DNA為模板,采用半定量RT-PCR方法獲得FABP4、ADIPOQ和CFD基因在阿勒泰羊不同組織中的表達(dá)譜。4、應(yīng)用實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)了FABP4、ADIPOQ和CFD基因在阿勒泰羊饑餓與非饑餓模型尾脂組織中的表達(dá)變化情況。結(jié)果:1、通過(guò)2周逐步限飼和4周完全禁食(期間自由飲水)后,饑餓模型阿勒泰羊精神上表現(xiàn)出焦躁不安、采食欲望強(qiáng)烈;其臀尾表現(xiàn)出“癟瘦”表型,臀尾部皮膚松弛、軟塌,臀部尾脂體積減小了1/3以上;體重顯著下降;尾脂脂肪細(xì)胞體積變小,呈扁平的梭形。2、克隆得到綿羊FABP4、ADIPOQ和CFD基因完整的CDS序列,分別編碼132、239、261個(gè)氨基酸。綿羊FABP4核苷酸序列及氨基酸序列與山羊、牛、牦牛、馬鹿等反芻動(dòng)物對(duì)應(yīng)序列的同源性均高達(dá)90%以上;綿羊ADIPOQ基因的核苷酸序列與牛、小鼠、大鼠、野豬、人等對(duì)應(yīng)序列的同源性在75%以上,其中與牛對(duì)應(yīng)序列的同源性高達(dá)85%以上;綿羊CFD基因核苷酸序列及其編碼的氨基酸序列與反芻動(dòng)物牛的同源性均高達(dá)96.9%以上。3、半定量RT-PCR檢測(cè)結(jié)果表明,FABP4基因在阿勒泰羊心臟、肺臟、肌肉、腸系膜脂肪、尾脂組織中均有表達(dá),但在腸系膜脂肪和尾脂中高豐度表達(dá);ADIPOQ基因在尾脂組織中特異性高表達(dá);CFD基因在心臟、肝臟、脾臟、肺臟、肌肉、尾脂中均表達(dá),但在尾脂中高豐度表達(dá)。4、實(shí)時(shí)熒光定量PCR結(jié)果表明,阿勒泰羊FABP4基因在非饑餓組尾脂中的表達(dá)量略高于饑餓組尾脂,但二者差異不顯著(P0.05);ADIPOQ、CFD基因則結(jié)果相反,阿勒泰羊ADIPOQ和CFD基因在饑餓組尾脂中的表達(dá)量顯著高于在饑餓組尾脂中的表達(dá)(P0.01)。結(jié)論:上述結(jié)果表明,FABP4、ADIPOQ、CFD基因在阿勒泰羊尾脂沉積與代謝過(guò)程中具有一定的調(diào)控作用,可以作為哺乳動(dòng)物脂肪組織發(fā)育調(diào)控過(guò)程中重要的候選功能基因。
[Abstract]:Objective: Altay Sheep is a kind of fat-buttock sheep breed. Its unique tail fat plays an important role in maintaining normal metabolism and body temperature. It is an ideal model to study the mechanism of tail fat deposition and metabolism in sheep, mainly through the regulation of tail fat deposition and metabolism to adapt to bad environment to maintain normal metabolism and body temperature. Recent studies have shown that FABP4ADIPOQ and CFD genes are involved in glucose and fatty acid metabolism and adipocyte differentiation. In this paper, the effect of FABP4 ADIPOQN CFD gene on fat deposition and metabolism of tail fat in Altay sheep was studied. Methods: 1, the model of starvation and non-starvation in Altay sheep. Two extreme states of tail fat deposition and metabolism were simulated in Altay sheep under natural physiological conditions, that is, fat deposition during summer and autumn, and catabolism of tail fat in spring and winter when forage was scarce. The animal model of tail lipid deposition and metabolism under starvation and non-starvation in Altay sheep was constructed. The complete CDS sequence of FABP4ADIPOQ and CFD gene was cloned by RT-PCR from the cDNAs of Altay sheep tail fat tissue. The CDS region of FABP4ADIPOQ and CFD gene was cloned from the heart and liver of Altay sheep. The main tissues of spleen, lung, kidney, muscle, mesenteric fat, tail fat, etc. The expression profiles of FABP4AADIPOQ and CFD genes in different tissues of Altay sheep were obtained by semi-quantitative RT-PCR. The expression of FABP4AADIPOQ and CFD genes in tail fat tissue of Altay sheep model of starvation and non-starvation was detected by real-time fluorescent quantitative PCR. The results showed that after 2 weeks of diet restriction and 4 weeks of complete fasting (free drinking), Altay sheep showed mental agitation and strong appetite for food intake, and its buttocks and tails showed "punk" phenotype and loose skin of buttocks and tails. After soft collapse, the volume of fat in the buttocks decreased by more than one third, the weight of fat cells decreased significantly, and the fat cells of tail fat became smaller, showing flattened fusiform. The complete CDS sequences of FABP4ADIPOQ and CFD genes were cloned and encoded 132239261 amino acids, respectively. The nucleotide sequence and amino acid sequence of sheep FABP4 were more than 90% homology with goat, cow, yak, red deer and other ruminants, and the nucleotide sequence of sheep ADIPOQ gene was higher than that of cattle, mice, rats, wild boar, etc. The homology of human equivalent sequence is more than 75%, and the homology with cattle is more than 85%. The nucleotide sequence of sheep CFD gene and the amino acid sequence encoded by CFD gene were above 96.9%. The results of semi-quantitative RT-PCR showed that FABP4 gene was found in the heart, lung, muscle and mesenteric fat of Altay sheep. The high abundance of ADIPOQ gene was expressed in mesenteric fat and tail fat, and the expression of CFD gene in heart, liver, spleen, lung, muscle and tail fat was also found in heart, liver, spleen, lung, muscle and tail fat. The results of real-time fluorescence quantitative PCR showed that the expression of FABP4 gene in non-starvation group was slightly higher than that in hungry group, but there was no significant difference between them (P0.05). The expression of ADIPOQ and CFD genes in the tail fat of the hungry group was significantly higher than that in the starved group (P0.01). Conclusion: these results suggest that FABP4 ADIPOQN CFD gene may play an important role in the deposition and metabolism of tail fat in Altay, and it can be used as an important candidate gene in the regulation of adipose tissue development in mammals.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S826

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